The growth rate of tubercle bacilli from South Indian and British patients

CULTURES of tubercle bacilli from Indian patients have been shown to be, on average, less virulent in the guinea-pig and to have a wider range of virulence than cultures obtained from British patients (Frimodt-Moller, Mathew and Barton, 1956 ; Mitchison et al., 1960 ; Bhatia et al., 1961). In the study of Bhatia et al. (loc. cit.) about one-third of the Indian cultures were as virulent as British cultures, the remainder being less virulent. In these studies the extent of disease in the organs of the guinea-pig was scored at intervals after the intramuscular injection of the organisms. In consequence, the measure of virulence was based upon the rate of development of the lesions and, by inference, the rate of multiplication of the bacilli in the organs. It was, therefore, considered of interest to compare the growth rates in vitro of Indian and British cultures of tubercle bacilli

Non-identity of reaction centres for pyrophosphatase and toxic actions of cardiotoxin II: the status of cardiotoxin II as a metalloprotein

Cardiotoxin II of the Indian cobra (Naja naja) contains approximately four Mg2+ per mol. Complete demetallation of the toxin is achieved by three cycles of treatment with ethylenediamine tetraacetate and gel filtration. Reconstitution of toxin by treatment of the apo-protein with Mg2+ restores metal content and inorganic pyrophosphatase activity only to the extent of two atoms/mol and 65%, respectively. Use of Mg (II)-EDTA in the reconstitution experiment yields restoration of half the original enzyme activity. Mg2+ is required for the inorganic pyrophosphatase action of the toxin. A definitive statement on the non-essentiality of Mg2+ for the lethal toxicity of the toxin is not possible at present, although experimental observations indicate that demetallated toxin is as toxic as the native toxin. Based on this and the differing sensitivities of the enzyme and toxic activities of the toxin to heat, it is suggested that the reaction centres in the toxin for the two activities are different and that the pyrophosphatase activity is not causally connected with the lethal toxicity of the toxin

Anomalous lateral diffusion in a viscous membrane surrounded by viscoelastic media

We investigate the lateral dynamics in a purely viscous lipid membrane surrounded by viscoelastic media such as polymeric solutions. We first obtain the generalized frequency-dependent mobility tensor and focus on the case when the solvent is sandwiched by hard walls. Due to the viscoelasticity of the solvent, the mean square displacement of a disk embedded in the membrane exhibits an anomalous diffusion. An useful relation which connects the mean square displacement and the solvent modulus is provided. We also calculate the cross-correlation of the particle displacements which can be applied for two-particle tracking experiments.Comment: 6 pages, 2 figure

Effect of N-bromosuccinimide-modification of tyrosine side chains of cardiotoxin II of the Indian cobra on biological activity

The essential role of tyrosine residue(s) of cardiotoxin II in the biological activity of the toxin was evaluated using N-bromosuccinimide. N-bromosuccinimide effected oxidation of the tyrosine residues in cardiotoxin II with enhancement in absorbance at 260 nm. The influence of various solvent media such as acetate-formate buffer (pH 4.0), 0.01 N H2SO4 (pH 2.0) and Tris-HCl buffer (pH 8.5) on oxidation of tyrosine residues was exa mined. In comparison with 0.01 N H2S O4, acetate-formate buffer could prevent secondary oxidations as revealed by lower consumption of oxidant, N-bromosuccinimide, to achieve oxidation. In Tris-HCl buffer oxidation of tyrosine did not take place effectively. N-iodo-succinimide caused only limited oxidation as evident from minor increase in absorbance at 260 nm. N-chlorosuccinimide was completely ineffective. Oxidation of cardiotoxin II with 3.75 equivalents of N-bromosuccinimide tyrosine residue led to complete loss of lethal activity. However, the derivative retained the ability to protect bacterial protoplasts from lysis in solutions of low tonicity. Unlike cardiotoxin II oxidized with N-chlorosuccinimide (50 equivalents/mol of toxin) which retained lethal activity as well as the ability to protect protoplasts from lysis, performic acid-oxidized toxin had lost both the activities

The action of sulpeiuric acid on gliadin: with special reference to the N-Peptpdyl→O-Peptpdyl bond rearrangement

Treatment of gliadin with sulphuric acid transposes peptide bonds of serine from the amino to the hydroxyl group. Maximum transposition, 60-70% of the theoretical, occurs when the protein is treated with anhydrous sulphuric acid at 0° C. for 35 hr. No rearrangement was detected at threonine residues. Examination of the peptide material, obtained from the rearranged protein by Elliott's degradation method, indicates apparent "homogeneity". In an alternative scheme for the degradation, nitrous acid deamination of free amino groups was used. The resulting loss in serine content of the protein is direct evidence for the acyl migration of peptide bonds. Incorporation of sulphur and partial disappearance of several amino acids accompany the sulphuric acid treatment. The occurrence of these secondary reactions imposes limitations on the use of sulphuric acid as a reagent for the specific fission of peptide bonds

The terminal amino acids of wheat gliadin

Wheat gliadin has been found by two different methods to contain three N-terminal histidine residues for each molecular weight of 27,000. Trace amounts of N-terminal aspartic acid, glutamic acid, alanine, valine, and serine were also detected in the preparation used. Hydrolysis in boiling hydrochloric acid partially destroyed the di-2,4-dinitrophenyl derivative of histidine. Losses of from 5% to 25% occurred depending upon the time and conditions of hydrolysis. Carboxypeptidase did not release free amino acids from wheat gliadin but qualitative evidence for the occurrence of C-terminal glutamic acid and C-terminal "leucine" was obtained

Probing the mechanism of electron capture and electron transfer dissociation using tags with variable electron affinity

Electron capture dissociation (ECD) and electron transfer dissociation (ETD) of doubly protonated electron affinity (EA)-tuned peptides were studied to further illuminate the mechanism of these processes. The model peptide FQpSEEQQQTEDELQDK, containing a phosphoserine residue, was converted to EA-tuned peptides via β-elimination and Michael addition of various thiol compounds. These include propanyl, benzyl, 4-cyanobenzyl, perfluorobenzyl, 3,5-dicyanobenzyl, 3-nitrobenzyl, and 3,5-dinitrobenzyl structural moieties, having a range of EA from −1.15 to +1.65 eV, excluding the propanyl group. Typical ECD or ETD backbone fragmentations are completely inhibited in peptides with substituent tags having EA over 1.00 eV, which are referred to as electron predators in this work. Nearly identical rates of electron capture by the dications substituted by the benzyl (EA = −1.15 eV) and 3-nitrobenzyl (EA = 1.00 eV) moieties are observed, which indicates the similarity of electron capture cross sections for the two derivatized peptides. This observation leads to the inference that electron capture kinetics are governed by the long-range electron−dication interaction and are not affected by side chain derivatives with positive EA. Once an electron is captured to high-n Rydberg states, however, through-space or through-bond electron transfer to the EA-tuning tags or low-n Rydberg states via potential curve crossing occurs in competition with transfer to the amide π* orbital. The energetics of these processes are evaluated using time-dependent density functional theory with a series of reduced model systems. The intramolecular electron transfer process is modulated by structure-dependent hydrogen bonds and is heavily affected by the presence and type of electron-withdrawing groups in the EA-tuning tag. The anion radicals formed by electron predators have high proton affinities (approximately 1400 kJ/mol for the 3-nitrobenzyl anion radical) in comparison to other basic sites in the model peptide dication, facilitating exothermic proton transfer from one of the two sites of protonation. This interrupts the normal sequence of events in ECD or ETD, leading to backbone fragmentation by forming a stable radical intermediate. The implications which these results have for previously proposed ECD and ETD mechanisms are discussed

Hydrodynamic coupling between two fluid membranes

The coupled in-plane diffusion dynamics between point-particles embedded in stacked fluid membranes are investigated. We calculate the contributions to the coupling longitudinal and transverse diffusion coefficients due to particle motion within the different as well as the same membranes. The stacked geometry leads to a hydrodynamic coupling between the two membranes.Comment: 9 Pages, 5 figures. Accepted for publication in J. Phys.: Condens. Matte

Effects of an embedding bulk fluid on phase separation dynamics in a thin liquid film

Using dissipative particle dynamics simulations, we study the effects of an embedding bulk fluid on the phase separation dynamics in a thin planar liquid film. The domain growth exponent is altered from 2D to 3D behavior upon the addition of a bulk fluid, even though the phase separation occurs in 2D geometry. Correlated diffusion measurements in the film show that the presence of bulk fluid changes the nature of the longitudinal coupling diffusion coefficient from logarithmic to algebraic dependence of 1/s, where s is the distance between the two particles. This result, along with the scaling exponents, suggests that the phase separation takes place through the Brownian coagulation process.Comment: 6 pages, 5 figures. Accepted for publication in Europhys. Let